Table 1.
Primers and TagMan-MGB probes for real-time assays targeting F. tularensis.
| Assay | Region in Genome of SCHU S4 a |
Sequences of Forward and Reverse Primers b and Probes c | Primer (µM) d | MGB e | Probe (µM) f | Tm °C g |
|---|---|---|---|---|---|---|
| Ft-sp. FTT0376 | 377718–377824 | F CCATATCACTGGCTTTGCTAGACTAGT R TGTTGGCAAAAGCTAAAGAGTCTAAA FAM-AAATTATAAAACCAAACCCAGACCTTCAAACCACA |
0.9 | no | 0.2 | 60 |
| Ft-sp.3Pan | 8967–9049 | F TTTACACCCGTCTCCGTTAGT R CTCTTAAGGATGCAATTTGGGATT FAM-AAGAGGCAAAGCTGGAATTACACTCTCTC |
0.9 | no | 0.2 | 60 |
| Ft-sp. FTS_0772 | 1187646–1187807 | F CAAGGTAAAGAAATTAAGAGTAGTAAAGTTGAATTC R ATTTAATCTAGTATTATCAATTGGGTAAAAAGGTA FAM-GATGTGGCAACAACTGAAAT |
0.9 | yes | 0.2 | 63 |
a Position coordinates of assay target within reference SchuS4 whole genome sequence (AJ749949.2); b F: forward primer, R: reverse primer; c 6-carboxyfluorescein (FAM) fluorophore and a 3= black hole quencher-1 (BHQ-1) quencher; d Primer concentration (μM) per reaction; e Minor Groove Binding moiety on probe; f Probe concentration (μM) per reaction; g Real-time PCR annealing temperature.